The Primary Study On Extraction And Molecular Characteristics Of Collagen From Skin Of Scomberomorus Niphonius

This article was to research on collagen’s extract method,physico-chemical property, depuration and molecule characteristics from skin of Scomberomorus niphonius. This article in order to lays the foundation in utilize and develop and supply an theory in it. The major results were as following:1. This text was adopted acid and enzymic to extract collagen. The result of single factor test showed that the organic was best medium than other acid and pepsin was the best hydrolase than other different enzyme. The results of L9(34) orthogonal test abstracted collagen with acid showed the highest extraction rate was23.51%, the best extraction condition was like that:0.2mol/L citric acid; the ratio of material and fluid was1:30, the swelling time was48h. The results of L9(34) orthogonal test abstracted collagen with pepsin showed the highest extraction rate was35.73%, the best extraction condition was like that:3%pepsin(w/v); the ratio of material and fluid was1:30; the reaction time was6h; the temperature was30℃. The results of L9(34) orthogonal test abstracted collagen with pepsin and supersonic wave showed the highest extraction rate was82.42%, the best extraction condition was like that:3%pepsin(w/v); raw material was1:30; swelling time was3h; the temperature was30℃. We use SDS-PAGE electrophoregram to compare relative molecular weight, and we know the production extracted by pepsin and supersonic wave was smaller than production extracted by acid.2. Spectral characteristic of collagen from skin of Scomberomorus niphonius. Collagen had striking absorption peak in227nm,230nm and non-assimilation in260nm, because of collagen’s amino acid composition hardly exist aromatic amino acid which had conjugated double bond. FT-IR showed collagen had striking absorption peak in3427cm-1,1651cm-1,1544cm-1and1240cm-1, those regions were amides A, amides Ⅰ, amides Ⅱ and amides Ⅲ, so we can said that the collagen keep its triple helical structure well.3. We use DEAE-52ion exchange chromatography to depurate ASC from skin of Scomberomorus niphonius. The result was that we got two non-resolved peaks, the fist one was proteoglycan and second was collagen. The experiment condition was like that:toplimit of saltness was100mmol/LNaCl, flow rate was30mL/h, the starting saltness was13mmol/L, the frist eluting peak’ saltness was22.25mmol/L, the second eluting peak’ saltness was30mmol/L; ending saltness38mmol/L4. Molecule characteristic of collagen from skin of Scomberomorus niphonius. The result of assaying collagen’s amino acid composition showed that it was rich in Gly(18.4%), Pro(8.51%), Ala(8.31%), Glu(7.77%), Arg(6.85%) were lower, and could not find Tyr. We could know collagen’s type, compose of chain, relative molecular weight by SDS-PAGE. The SDS-PAGE electropherogram showed that the collagen contained chains of α1,α2, β and γ, the molecular weight was127kDa,115kDa,278kDa,343kDa, and then this ASC was classified as type I collagen.